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322 PHL Lab # 1 Anti-inflammatory Drugs 2/24/2011 Samiah AL Shehri 1 Inflammation Definition: It is a protective response against injury which can be chemicals, mechanical, infectious, or immunological. 2/24/2011 Samiah AL Shehri 2 Anti- inflammatory Drugs Signs of inflammation Erythema )redness) Oedema Tenderness & pain Heat vasodilation vasodilation main pain mediator is Leukotriene Chemical mediators of inflammation Bradykinin Leukotriene Prostaglandin Histamine 2/24/2011 Pain VD + blood flow (heat+redness+oedema) odema + VD Samiah AL Shehri 3 Cellular events during inflammation 1. Acute stage: Vasodilatation and increase vascular permeability due to the release of histamine & 5HT leading to oedema. 2. Exudative stage or delay or subacute: Migration of the leukocytes to the site of inflammation. 3. Repair stage or chronic proliferation phase: Proliferation of fibroblast or formation of new connective tissue (fibrosis). 2/24/2011 Samiah AL Shehri 4 Classification of the Anti-inflammator Drug (AIDs) Type SAIDs NSAIDs Examples Hydrocortisone, dexamethasone Aspirin, Paracetamol, & Indomethacin MOA Induce protein called lipocortin which cause inhibition of phospholipase A2 Inhibit cyclooxygenase enzyme leading to decrease prostaglandin biosynthesis (PGE1 & PGI2) 2/24/2011 Samiah AL Shehri 5 Site of Action of The Anti-inflammatory Drugs Phospholipids (-) SAIDs (+) Phospholipase A2 Arachidonic Acid Lipooxyge nase (-) NSAID Leukotrienes (+) COX PG PAF (platelet aggregating factors) 2/24/2011 Samiah AL Shehri 6 Site of Action of The Anti-inflammatory Drugs N.B. Aspirin is contraindicated for asthemitic patients …? because it inhibits the COX enzyme shifting the reaction to the lipooxygenase pathway leading to increase the leukotrienes production which will cause bronchocostrictions. 2/24/2011 Samiah AL Shehri 8 Screening Methods for The Antiinflammatory Drugs 1. Rat - Paw Oedema Method (Acute stage) 2. Granulation Pouch Method (Exudative stage) 3. Adjuvant induce arthritis. 2/24/2011 Samiah AL Shehri 9 Granulation Pouch Method Used to measure the activity of anti-inflammatory drugs by induction of granuloma causing exudative proliferative effect in two main groups control & treated . Animal: Rats. Irritant substance: croton oil or carragenin . Drug used: anti-inflammatory drugs . 2/24/2011 Samiah AL Shehri 10 3-Adjuvant induced arthritis Animal: Rats. Irritant substance: tubercle bacilli . Drug used: anti-inflammatory drugs . 2/24/2011 Samiah AL Shehri 11 Rat - Paw Oedema Method Animal: 4 Rats ( 2 control and 2 treated ) . Irritant substance: Carragenin (2%) . Drug used: Indomethacin (2 mg/kg) . Procedure : For the control gp inject 0.1 ml S.C into planter or sub planter tissue of the rat hind paw of one foot, then left the rat for 3-4 hrs for development of inflammation. 2/24/2011 Samiah AL Shehri 12 Rat - Paw Oedema Method For the treated gp inject the indomethacin (2 mg/kg) I.P 40 -45 min. before, then inject the carragenin S.C and left the rat for oedema development . Measure the oedema produced by both control and treated gp and calculate the % reduction of oedema by indomethacin . 2/24/2011 Samiah AL Shehri 13 RESULTS 2/24/2011 Results Control Groups Treated Groups Position 1 2 Mean Right ------A Right ------C Samiah AL Shehri Left ------B Left ------D 14 CALCULATIONS 1. * % of oedema produced by carragenin alone = A – B/B ×100 = X % 2. * % of oedema produced by carragenin after treatment = C – D/D ×100= 3. 4. Y% * % Reduction of oedema by indomethacin = X – Y = % NOTE: A, C = Mean right for control B, D = Mean left for control 2/24/2011 Samiah AL Shehri 15 2/24/2011 Samiah AL Shehri 16